AIM: To investigate the expression of matrix metalloproteinases (MMPs), membrane type MMPs (MT-MMPs) and their inhibitors (TIMPs) in human primary cultured prostatic cells and malignant prostate cell lines.METHODS: Reverse transcription-polymerase chain reaction-based measurements of the mRNA levels of MMP-2, MMP-7, MMP-9, MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 in relation to the house-keeping gene glyceraldehydes phosphate dehydrogenase were performed in cancerous and non-cancerous prostatic tissue samples, in primary cell cultures of epithelial cells, in both fibroblasts and smooth-muscle cells as stromal cells, and in the human malignant prostatic cell lines DU-145, LNCaP and PC-3.RESULTS: MMP-2 was mainly expressed in stromal cells. MMP-7 and MMP-9 showed their highest values in epithelial cells. MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 were found both in stromal and in epithelial cells, but there were some differences between the expression in fibroblasts and smooth-muscle cells. Different expression was also observed between the cells deriving from the primary cell cultures, the benign cell line BPH-1, and the malignant cell lines LNCaP, DU-145, and PC-3.CONCLUSION: These results concerning different expression of MMPs and TIMPs in cells from prostatic tissue suggest that a better insight into changes observed in prostatic tissue needs studies on cells cultured from the tissue.

Asthma ; diagnosis ; Child ; Humans ; Perception

OBJECTIVETo investigate the effect of hypoxia on the human pulmonary artery smooth muscle cells two pore domain potassium channels TASK-1 and the regulation of non-receptor tyrosine kinase c-Src in this process.

METHODSThe cultured human pulmonary artery smooth muscle cells (hPASMCs) were divided into: normal group, hypoxia 30 minute group, hypoxia 6 hours group and hypoxia 48 hour group, and hypoxia 48 hour + PP2 group, hypoxia 48 hour + PP3 group, hypoxia 48 hour + bpV group. Flow cytometry was used to analyze the cell cycle, RT-PCR and Western blot technique were carried out to detect the expression changes of TASK-1 mRNA and protein in different groups.

RESULTS(1) Cell Cycle Show: Compared with normal control group, with prolonged hypoxia, the percentages of hPASMCs in S phases of cell cycle were increased. While compared with hypoxia 48 hour group, the percentages of hypoxia 48 hour + PP2 group hPASMCs in S phases of cell cycle were decreased. The expression of TASK-1 mRNA on hPASMCs in acute hypoxia 6 hour group was increased, while the expression of TASK-1 protein on hPASMCs in the acute and chronic hypoxia group was decreased, and the expression of TASK-1 mRNA on hPASMCs in the chronic hypoxia group was decreased; After pre-incubation of a potent and selective inhibitor of the Src family of protein tyrosine kinases PP2, the expression of TASK-1 mRNA and protein in hypoxia 48 hour group was increased, however after pre-incubation of the inhibitor of the Src family of protein tyrosine phosphatase bpV, the expression of TASK-1 protein in hypoxia 48 hour group was decreased.

CONCLUSIONHypoxia promotes human pulmonary artery smooth muscle cell proliferation, and non-receptor tyrosine kinase c-Src may participate in the expression of two pore domain potassium channels TASK-1 regulated by hypoxia. Therefore, we hypothesized that TASK-1 channels and c-Src participatein the acute and chronic hypoxic human pulmonary vasoconstriction.

Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology ; Nerve Tissue Proteins ; metabolism ; Potassium Channels, Tandem Pore Domain ; metabolism ; Pulmonary Artery ; cytology ; RNA, Messenger ; Vasoconstriction ; src-Family Kinases ; metabolism

Objective To investigate homografting vascular endothelial progenitor cells(EPCs)for preventing restenosis formation of carotid artery in New Zealand white rabbit models.Methods EPCs of New Zealand white rabbits were isolated,confirmed and expanded though the injured carotid arterial endothelium of rabbit model induced by dilatation with a 2.5 F balloon;and then EPCs were transplanted into the injured endothelium of the cells transplantation group(n=13,3 of them were transplanted with fluorencently-labeled- EPCs),while equal volume of saline without EPCs was injected into the injured endothelium in the control group(n=8).Histopathology was performed at 4 days after transplantation for the 2 rabbits,with fluorencently-labeled-EPCs.All of the rest remained rabbits were killed 4 weeks later for histological examinations.Results The histopathological slides showed that the fluorescence-positive expression existed in the injured endothelium 4 days after transplantation.At 4 weeks after the EPCs transplantation,there were less restenosis and less vascular wall thickening in the rabbits of cells transplantation group than those of the control group(P＜0.01).Conclusion The local interventional homografting heterogeneous endothelial progenitor cells can prevent restenosis after the carotid artery angioplasty in New Zealand White rabbit model. (J Intervent Radiol,2007,16:95-98)

OBJECTIVEPrepare konjac glucomannan-hydroxypropyl methyl cellulose (HPMC) compression coated tablets and study the effects of the formulation, technics and in vitro dissolution condition on drug release behavior to elevate the colon-specific effects of preparation.

METHODBerberine hydrochloride core tablets were prepared by wet granulation technique and konjac glucomannan-HPMC mixture as the coating layer were used with compression coated technique. The effects of the formulation and technics on drug release behavior were investigated by dissolution test. The erosion of coat layer during dissolution test was investigated.

RESULTDrug almost not released in dissolution medium stimulating gastric and intestinal condition, and released completely by coating layer erosion and rupture by enzyme in stimulating colonic condition. Drug release decreased with decreasing the ratio of konjac glucomannan-HPMC and increasing coat weight (P < 0.05), compression force was not found to be a significant factor on drug release. Drug release increased with increasing the concentration of beta-mannase in dissolution medium (P < 0.05), rotation speed has no effect on drug release. The release of drug was correlative with erosion of coat layer. The mechanism of drug release were diffusion and erosion.

CONCLUSIONThe konjac glucomannan-HPMC compression coated tablets was a promising delivery system for drugs to be delivered to the colon.

Administration, Oral ; Amorphophallus ; chemistry ; Berberine ; administration & dosage ; chemistry ; pharmacokinetics ; Colon ; metabolism ; Drug Compounding ; methods ; Drug Delivery Systems ; Hypromellose Derivatives ; Mannans ; chemistry ; isolation & purification ; Methylcellulose ; analogs & derivatives ; chemistry ; Plants, Medicinal ; chemistry ; Tablets, Enteric-Coated

BACKGROUNDActinomycosis is a rare indolent infectious disease caused by Actinomyces. Although pulmonary actinomycosis is thought to be more prevalent in developing countries, data from developing countries are scanty. This study was to reveal the current situation of pulmonary actinomycosis in developing countries and the difference from that in developed countries.

METHODSPatients fulfilling the inclusion criteria for pulmonary actinomycosis from Peking Union Medical College Hospital in China between January 2003 and December 2014 were retrospectively analyzed. Baseline characteristics, clinical symptoms, underlying diseases, diagnostic methods, pulmonary function test results, chest computed tomography (CT) tests, fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) tests, initial diagnosis, treatment and prognosis were retrieved from medical records and analyzed.

RESULTSTwenty-six patients were included in this study (mean age 52.0 + 13.1 years). The ratio of male to female was 1.17:1. Most common clinical symptoms were cough (15/26), sputum (12/26) and hemoptysis (12/26). Chest CT findings presented as masses (13/26), nodules (10/26) and infiltrates (3/26). FDG-PET had an increased standardized uptake value and 4/6 patients were misdiagnosed as malignancy. Many kinds of antibiotics were used in the treatment of pulmonary actonomycosis and all got favorable results. Five patients receiving complete resection of the lesion were cured without postoperative use of antibiotic.

CONCLUSIONSPulmonary actinomycosis is a rare disease even in developing countries, and both misdiagnosis and missed diagnosis are common. FDG-PET seems useless in the differential diagnosis, and complete resection of the pulmonary lesion without postoperative antibiotic therapy might be enough to achieve cure.

Actinomycosis ; diagnosis ; diagnostic imaging ; metabolism ; Adult ; Aged ; China ; Female ; Fluorodeoxyglucose F18 ; Humans ; Lung Diseases ; diagnosis ; diagnostic imaging ; metabolism ; Male ; Middle Aged ; Positron-Emission Tomography ; Radiography ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo establish a new method based DNA chip technique for detecting HCV genotypes.

METHODSGenotyping probes were designed according to the sequence of HCV 5' NCR to generate DNA chip. The probes on DNA chip contains 5 major genotypes and 8 subtypes. The DNA fragment amplified by labeling Cy5 fluorescence was hybridized with DNA chip.

RESULTSFifty-five out of 65 isolates detected by DNA chip belonged to 1b- DNA sequencing of form a part of the isolates was used as the control. The results of both were completely consistent.

CONCLUSIONThe method is simple and rapid with high specificity and sensitivity. It can be applied in detection of HCV RNA and genotypes.

5' Untranslated Regions ; genetics ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; RNA, Viral ; blood ; Sensitivity and Specificity

OBJECTIVETo evaluate the expression of microRNAs and reveal the regulatory mechanism of miRNA-708 in pediatric common acute lymphoblastic leukemia (ALL) (common-ALL).

METHODSThe expressions of microRNAs in common-ALL patients were detected by microarrays in 3 pediatric common-ALL samples, and then verified by stem-loop quantitative RT-PCR in 34 common-ALL samples. The target genes of miR-708 were found by bioinformatics software, and verified by dual-luciferases reporter assay, RT-PCR and Western blot.

RESULTSCompared to normal bone marrow samples, of all the 2006 detected miRNAs, the expression of miR-708, miR-181b and miR-210 were 16.886 ± 16.854, 5.710 ± 4.652, and 9.789 ± 1.178, retrospectively, being significantly up-regulated expressed than those in normal control (1.872 ± 0.339, 1.276 ± 0.531 and 1.005 ± 0.080, retrospectively) (P < 0.05), while miR-27b and miR-345 were the two most down-regulated ones (0.524 ± 0.085 and 0.675 ± 0.086, retrospectively) (normal control: 1.123 ± 0.066 and 1.204 ± 0.140, retrospectively) (P < 0.05). And the expression of miR-708 and miR-181b were significantly correlated with the clinical types in common-ALL. In high risk common-ALL, miR-708 and miR-181b were much higher than in standard and middle risk common-ALL (P < 0.05). The further verification research in 293 cell line showed that miR-708 decreased the expression level of its target genes CNTFR, NNAT and GNG12 by combining with 3'-UTR of the 3 genes, moreover, miR-708 combined with CNTFR 3'-UTR in 394 ∼ 400 bp sequence region.

CONCLUSIONMicroRNAs plays an important regulatory role during the occurrence and development of the pediatric common-ALL and miR-708 is an important factor for high risk common-ALL.

Child ; Child, Preschool ; Female ; Gene Expression Profiling ; Humans ; Infant ; Male ; MicroRNAs ; genetics ; metabolism ; Microarray Analysis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

Objective To explore the association between polymorphisms of interferon-gamma gene intron 1 at position+874 (IFN-γ+874) gene and the susceptibility of HBV and/or HCV infection with different clinical outcomes. Methods IFN-γ+874 gene SNP were detected in 277 subjects including 79 chronic HBV/HCV coinfections,69 individuals only with HBV infection,55 individuals only with HCV infection and 74 controls,by sequence specific primers-PCR (SSP-PCR). Hepatocellular injury as suggested by alanine aminotransferase (ALT) was detected by Beckman LX-20. The status of viral particles in serum was determined by RT-nPCR. The possible association of the polymorphism of IFN-γ+874 with the susceptibility of HBV and/or HCV infection and the outcome of these infections were analyzed. Results (1) IFN-γ+874 AA frequency in individuals with chronic HBV,HCV,HBV/HCV coinfections were significant higher than that in controls (X~2=16.15,P=0.01); OR (95% CI) of IFNγ+874 AA in chronic infection with HBV,HCV,HBV/HCV coinfections appeared to be 2.70 (1.24-5.92),3.22 (1.43-7.25) and 4.02 (1.88-8.55) compared with + 874 TA. No significant differences were found among HBV,HCV,HBV/HCV coinfections (X~2=1.97,P=0.73). There were no significant association of IFN-γ +874 A/T allele frequency with HBV and/or with HCV infection (X~2=4.87,P=0.18). (2)The clinical outcomes of mild chronic hepatitis (CH),moderate/severe CH and cirrhosis with HBV and/or HCV infection were associated with IFN-γ+874 AA [X~2＝14.17,P＝0.03;OR＝3.09(1.51-6.33),3.85 (1.70-8.70),3.14 (1.08-9.17)]. No significant relationships were found between IFN-γ+874 A/T allele frequency and the clinical outcome of HBV/HCV infection (X~2＝6.07,P＝0.11). (3)There were no significant associations of IFN-γ+874 genotype/allele frequency with HCV duplication (X~2=2.36,P=0.31). (4) There were no significant associations of IFN-γ+874 genotype/allele frequency with abnormal ALT (X~2=0.15,P=0.93). Conclusion These results suggested that polymorphisms in the IFN-γ +874 had some influence on chronic HCV and/or HBV infection,and on the outcome of HCV and/or HBV infections. IFN-γ+874 AA genotype and T allele were possible risk to chronic HBV and/or HCV infections and to the outcomes of HBV and/or HCV infection. However,IFN-γ+874 TA genotype might serve as possible protective factors to them.

Objective To study the relationship between polymorphisms in interleukin-2gene at position-330 (IL-2-330) and the clinical outcome of hepatitis B virus (HBV) and/or hepatitis C virus (HCV) infection. Methods 277 subjects were recruited including 79 chronic HCV co-HBV infection, 55 chronic HCV infection, 69 chronic HBV infection and 74 controls. Single nucleotide polymorphisms of IL-2-330 was investigated by restricted fragment long polymorphism-PCR (RFLP-PCR). Hepatocellular injury, as revealed by alanine aminotransferase (ALT) was detected by Beckman LX-20 analyzer. The presence of hepatitis C viral particles in serum was determined by RT-nPCR. Results ( 1 ) IL-2-330 polymorphisms showed close association with persistent HBV and/or HCV infection. IL-2-330 TT was associated with an increased risk, but IL-2-330 GG with a reduced risk of persistent HBV and/or HCV infection (χ2=14.24, P=0.03 ) with ORs (95%CI) as 7.14(2.13-23.81 ), 3.46 (1.17-10.02) and 2.93 (1.15-7.46) respectively. However,IL-2-330 TT/GG did not significantly differ between patients with HBV and/or HCV infection (χ2=2.09, P=0.72). IL-2-330 T allele was associated with an increased risk, but the -330G allele was associated with a reduced risk of chronic HBV/HCV infection (χ2=12.33,P=0.01),with ORs (95% CI) as 2.26 (1.39-3.69) , 1.82 ( 1.09-3.03 ) and 1.73 ( 1.10-2.73 ) respectively. (2) IL-2-330polymorphisms showed significant association with the outcome of HBV and HCV infection ( χ2=13.52, P=0.04). IL-2-330 TT was associated with an increased risk, but-330 GG with a reduced risk of mild CH, moderate/severe CH, and cirrhosis. The ORs (95%CI) appeared to be 3.33(1.75-6.32), 3.31 (1.75-6.26), 11.23 (3.09-40.76) respectively. IL-2-330 T allele was associated with an increased risk, but the -330 G allele was associated with a reduced risk of mild CH, moderate/severe CH and cirrhosis (χ2= 12.32, P=0.01 ), with ORs as 1.86(1.32-2.63), 1.71 (1.27-2.31) and 2.77(1.57-4.89) respectively. (3) The polymorphisms of IL-2-330 showed no association with HCV RNA replication (χ2=0.83, P=0.66; χ2=0.20, P=0.66). The polymorphisms of IL-2-330 were not significantly associated with abnormal ALT ( χ2= 1.10, P=0.58; χ2=0.08, P=0.78). Conclusion These results suggested that IL-2-330 TT/T was associated with an increased risk, but IL-2-330GG/G was associated with reduced risk of persistent HBV and/or HCV infection, and with the development of mild CH,moderated/severe CH,and cirrhosis.